枸杞多糖的分离、纯化、鉴定及生物活性的研究

The Isolation,Purification,Identification and Study on Biological Activities of Lycium Barbarum Polysacchuride

构杞子经热水提取,乙醇沉淀,获得构杞粗多糖。后者经 Sevag 法、蛋白酶法和透析纯化得到枸杞多糖(LBP)。LBP 进一步经 Sephadex G-50分子筛层析分离,可分为LBP_1(分子量>10000)和 LBP_2(分子量<10000)两种成分。LBP_1和 LBP_2用 DEAE-DE32离子交换层析去除色素,PAGE 电泳和 Brandford 法测蛋白质含量仅测到微量蛋白质(<5μg 蛋白/mg 糖)。经岛津 UV-260扫描,LBP、LBP_1、LBP_2未见核酸(260nm)和蛋白质(280nm)特征吸收峰,红外光谱分析表明有典型多糖吸收峰。分子量测定 LBP_1、LBP_2分别为35400、9900。体内外试验证明 LBP、BLP_1和 LBP_2对小鼠脾脏 T 淋巴细胞的功能呈双相调节,可明显促进 ConA 诱导的淋巴细胞增殖反应。抗体形成细胞试验显示 LBP_2还可明显增强小鼠体液免疫功能;并具抗辐射损伤作用,能明显延长 BALB/C 小鼠的生存期。

Lycium Barbarum(LB) was extracted by hot water,precipated by ETOH to obtain crude polysaccharide of LB, which was then treated by the method of proteinase digestion,Sevag method and dialysis to obtain LBP.Polyacrylamide tested by electrophoresis showed no obvious protein band.The quantitative test of protein by Brandford method exhibited no protein existing (5μg protein/mg polysaeeharide).The LBP was further purified by Sephadex G-50,two components were isolated:LBP1 (MW>10000) and LBP2 (MW<10000).Pigment was removed from LBP1 and LBP2 by DEAE- DE32 ion- exchange chromatography.Ultraviolet spectra analysis displayed no protein or nucleic acid.Infrared spectra analysis of LBP,LBP1 and LBP2 showed that there was characteristic polysaccharide absorption peak.The molecular weights of LBP1 and LBP2 were 35400 and 9900 respectively.Experiments in vitro and in vivo demonstrated that LBP, LDP1 and LBP2 had bi-direetion modulating effect on the proliferation of T lymphocytes of the mice spleens.They could increase the proliferation of ConA- stimulated T lymphocytes.LBP2 could significantly increased the number of mouse spleen antibody forming cell against SRBC,which indicated that LBP2 enhanced the function of humoral immunity of mice,it also had antagonistic effect on radiation-damage and significantly prolong the survival time of BALB/c mice.